Cytoskeletal polarization of T cells is regulated by an immunoreceptor tyrosine-based activation motif-dependent mechanism.
Binding of a T cell to an appropriate antigen-presenting cell (APC) induces the rapid reorientation of the T cell cytoskeleton and secretory apparatus towards the cell-cell contact site in a T cell antigen receptor (TCR) and peptide/major histocompatibility complex-dependent process. Such T cell polarization directs the delivery of cytokines and cytotoxic mediators towards the APC and contributes to the highly selective and specific action of effector T cells. To study the signaling pathways that regulate cytoskeletal rearrangements in T lymphocytes, we set up a conjugate formation assay using Jurkat T cells as effectors and cell-sized latex beads coated with various antibodies as artificial APCs. Here, we report that beads coated with antibodies specific for the TCR-CD3 complex were sufficient to induce T cell polarization towards the bead attachment site, as judged by reorientation of the microtubule-organizing center (MTOC) and localized actin polymerization. Thus, these cytoskeletal changes did not depend on activation of additional coreceptors. Moreover, single subunits of the TCR complex, namely TCR-zeta and CD3epsilon, were equally effective in inducing cytoskeletal polarization. However, mutagenesis of the immunoreceptor tyrosine-based activation motifs (ITAMs), present three times in TCR-zeta and once in CD3epsilon, revealed that the induction of cytoskeletal rearrangements required the presence of at least one intact ITAM. In agreement with this result, lack of functional Lck, the protein tyrosine kinase responsible for ITAM phosphorylation, abolished both MTOC reorientation and polarized actin polymerization. Both inhibitor and transient overexpression studies demonstrated that MTOC reorientation could occur in the absence of Ras activation. Our results suggest that APC-induced T cell polarization is a TCR-mediated event that is coupled to the TCR by the same signaling motif as TCR-induced gene activation, but diverges in its distal signaling requirements.
Lowin-Kropf
B
B
Department of Medicine, University of California, San Francisco, San Francisco, California 94143, USA.
Shapiro
V S
VS
Weiss
A
A
D000199:Actins / Q000737:chemistry / Q000378:metabolism; D000906:Antibodies / Q000502:physiology; D001665:Binding Sites / Q000235:genetics; D017252:CD3 Complex / Q000235:genetics; D018385:Centrosome / Q000502:physiology; D003599:Cytoskeleton / Q000737:chemistry / Q000502:physiology*; D016231:Fluorescence Polarization Immunoassay; D015870:Gene Expression / Q000235:genetics / Q000502:physiology; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology; D005799:Genes, Dominant / Q000235:genetics / Q000502:physiology; D011905:Genes, ras / Q000235:genetics; D006801:Humans; D019169:Jurkat Cells; D008565:Membrane Proteins / Q000235:genetics; D008863:Microspheres; D009154:Mutation / Q000235:genetics / Q000502:physiology; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology; D011971:Receptors, Immunologic / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000502:physiology; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D015533:Transcriptional Activation; D014443:Tyrosine / Q000502:physiology; D051746:ZAP-70 Protein-Tyrosine Kinase
D000199:Actins; D000906:Antibodies; D017252:CD3 Complex; D008565:Membrane Proteins; D011948:Receptors, Antigen, T-Cell; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins; C054979:antigen T cell receptor, zeta chain; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D051746:ZAP-70 Protein-Tyrosine Kinase; C497261:ZAP70 protein, human